Detection of Sodium Azide by Heteronucleus 14N NMR spectroscopy and - - PowerPoint PPT Presentation

Detection of Sodium Azide by Heteronucleus 14N NMR spectroscopy and binding to Fullerene C60

Tamar Chachibaia1,2, Manuel Martín Pastor3.

• 1. Department of Analytical Chemistry, Food Science and Nutrition, Faculty of Pharmacy,

University of Santiago de Compostela, Spain

• 2. Department of Public Health and Epidemiology, Faculty of Medicine, Iv.Javakhishvili

Tbilisi State University, Tbilisi, Georgia

• 3. Magnetic Resonance Unit, CACTUS, University of Santiago de Compostela, Spain

Presentation on March 9, 2015 in the Institute of Experimental Pharmacology & Toxicology Department of Biochemical Pharmacology Slovak Academy of Sciences Bratislava

Iv.Javakhishvili Tbilisi State University University of Santiago de Compostela

Aim

The aim of our study is to propose 14N NMR heteronucleus spectroscopy as valuable chemical analytical method for detection of sodium azide, which is used as a starting substance for the synthesis of many drugs and APIs, or investigational drugs and compounds and not only limited to these.

Sodium azide is acute poison similar to cyanide. Due to its attractive chemical and physical properties it is widely used in many spheres including automotive industry, medicine, pharmaco-chemistry and even everyday life. Detection of sodium azide becomes more demanding nowadays than several decades ago. We propose to use of 14N NMR spectra to detect and quantify sodium azide in aqueous solutions and extrapolate calibration results for real time detection of unknown

• concentrations. The results of this methodology relying

in measurement of 1D 14N NMR spectra at the lowest concentration of sodium azide aqueous solutions.

Quantitative NMR spectroscopy in pharmaceutical applications

Hazards and risks associated with Sodium Azide

1) Explosive – used in airbags and detonators. 2) Acute poison similar to cyanide. Inhibiting mitochondrial cytochrome C oxidase (CO) causing cerebral hypoxia and death; NaN3 is contributing to fast elaboration of nitric oxide (NO) with concomitant collapse. In human intake of 0.7–2 g (10 mg/kg) sodiun azide can lead to death within half an hour, and oral ingestion of lower doses (0.004–2 mg/kg) of NaN3 cause harm to human health, and chronic exposure to very low doses – dementia, e.g. at workplace area [[1]]. [1] S. Chang and S.H. Lamm. (2003). Human health effects of sodium azide exposure: a literature review and analysis. Int. J.

• Toxicol. 2222:175–186.

Spheres of application NMR spectroscopy for NaN3 detection

• Pharmaco-chemical analysis
• Occupational workplace monitoring
• Forensic tests
• Environmental safety
• Food and beverage quality control
• Security (detection of explosives)

1954s clinical study of sodium azide for its hypotensive effect

Black, M. M., B. W. Zweifach and F. D. Speer (1954) “Comparison of Sodium azide in Normotensive and Hypertensive Patients.” Exp Biol Med 85:11. Although was demonstrated lowering of arterial blood pressure by NaN3, but due to neuro degenerative deleterious side effect was not approved for clinical use.

NaN3 in the content of Sartans

NaN3 is widely used as starting molecule in the synthesis of Sartans, containing tetrazoles. [1] List of some sartans: Candesartan, Irbesartan, Losartan, Valsartan. While the pharmaceutical companies extensively apply qNMR in drug discovery and development they mostly use HPLC in routine quality analysis rather than qNMR. [2] Even though one- and two-dimensional NMR spectroscopy and qNMR are capable of the quality evaluation of drugs the number of applications in international pharmacopoeias, e.g. the European Pharmacopoeia (PhEur) and United States Pharmacopoeia (USP) is limited.

[1]Subramanian N., Babu V., Jeevan R., Radhakrishnan G. (2009). Matrix Elimination Ion Chromatography Method for Trace Level Azide Determination in Irbesartan Drug. J. of Chromatographic Science, Vol. 47, 529-533. [2] Santosh Kumar Bharti, Raja Roy (2012) Quantitative 1H NMR spectroscopy. Trends in Analytical Chemistry, 35, 5-25.

List of Sartans

14N and 15N

Nitrogen is a nucleus of considerable chemical and biological

• importance. However, despite its high isotopic abundance

(99.63%), 14N has always been a nucleus difficult to observe in

• NMR. It is a spin-1 nucleus.

15N is a spin-1/2 nucleus and thus can be studied with relatively high resolution even in the solid state, but it suffers from a low natural abundance (0.37%), which translates to a poor sensitivity. While the number of published 15N NMR papers is disproportionately small relative to the importance of nitrogen, studies of 14N isotope are even scarcer. [1] [1] O’Dell, L.A. (2011). Direct detection of nitrogen-14 in solid-state NMR spectroscopy. Progress in Nuclear Magnetic Resonance Spectroscopy, 59 (4), 295–318.

Sample preparation

A) co-axial insert tube - 100% nitromethane (CH3NO2) – 600 microliters B) Sample (5 different concentrations of NaN3 water solution (9:1 H20/D2) 100 mM, 50 mM, 25 mM, 10 mM, 4 mM C) Assembled for analysis

Sodium azide 100 mM

14N NMR

DRX-500, 300 K in 30 seconds with 64 scans N-=N+=N-

• USC. CACTUS. 19 Nov. 2013

DRX-500, 300 K

Sodium azide 100 mM

1 hour

15N NMR

Expected signals at 250 and 100 ppm are not seen

• USC. CACTUS. 19 Nov. 2013

15N NMR spectrum of sodium azide (1M) in D2O

http://chem.ch.huji.ac.il/nmr/techniques/1d/row2/n.html#n14properties

5 1 1 5 2 2 5 3 3 5 4 4 5 5 1 2 3 4 5 6 7 8 9 1 1 1

1 4N

N M R n

• rm

a liz e d in te g ra l (2 4 p p m ) Conc (mM)

E qu a tio n y = a + b *x W e igh t N

• W

e igh tin g R e sid u a l S u m

• f

S qu a re s 1 5 ,8 1 5 6 A d

• j. R
• S

qu a re ,9 9 7 2 9 V a lu e S ta n da rd E rro r C

• n

c In te rce p t

,27 2 5 1 1 ,0 9 88 6 S lo p e 2 ,05 9 5 8 ,0 4 80 2

1 2 3 4 5 6 7 8 9 1 2 3 4 5 6 7 8 9 1 1 1

Conc (mM)

1 4N

N M R n

• rm

a lize d in te g ra l (5 6 p p m )

E qu a tio n y = a + b *x W e igh t N

• W

e igh tin g R e sid u a l S u m

• f

S qu a re s 3 4 ,8 9 2 A d

• j. R
• S

qu a re ,9 9 4 2 V a lu e S ta n d a rd E rro r C

• n

c In te rce p t 1 ,6 7 6 3 6 1 ,5 8 8 5 5 S lo p e 1 ,2 1 7 2 9 ,0 4 2 2 2

CH3NO2 (ref.) NaN3 NaN3 100 mM 50 mM 25 mM 10 mM 4 mM

Properties of Fullerene C60

C60 is like any electron-deficient molecule can accept from 1 to 6 electrons and C60 is converted into anion. In the role of donors will serve external electrical charge, alkali metal ions or organic molecules. Like alkenes fullerene could be involved in the reaction of azide-alkyne cycloaddition, with the formation of triazole rings.

• A. R. Akhmetov, A. R. Tuktarov, U. M. Dzhemilev, I. R. Yarullin, and L. A. Gabidullina (2011). First example of

the interaction of fullerene C60 with hydrazoic acid. Russian Chemical Bulletin, International Edition, 60 (9), 1885—1887.

Results

The results demonstrate that there are changes in the chemical shift position and line-broadening related to the molar ratio NaN3:C60 in the sample (100:1). These results can be interpreted as binding interaction occurring between NaN3 and C60 molecules. As you will see in the attached figure, from the two 14N peaks of NaN3, the one that is more affected is the one that resonates at aprox. 56 ppm, which corresponds to two external nitrogen atoms.